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Updated: Jul 15, 2026
A Convenient and General Expression Platform for the Production of Secreted Proteins from Human Cells
Published on: July 31, 2012
[Steady expression and activity determination of recombinant signal peptide of mBD2 in SiHa cell]
Xiao-Li Wei1, Ming-Yuan Li, Hong Li
1Department of Microbiology, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China.
Objective:
To observe the expression pattern and effect of recombinant murine beta defensin 2 (rmBD2) on the proliferation of cell transfected with pcDNA3. 1 (+)/rmBD2.
Methods:
The recombinant plasmid pcDNA3. 1 (+)/rmBD2 was transferred into SiHa cells. The transfected SiHa cells were selected by G418 with 100 microg/mL for over 20 days. The steady expressions of rmBD2 protein and rmBD2 mRNA were detected by immunofluorescence and RT-PCR. The effects of rmBD2 on SiHa cell growth and reproduction were measured by MTT.
Results:
The SiHa cells which could stably express the rmBD2 protein were harvested with 100 microg/mL of G418. The rmBD2 protein expression was, by immunofluorescence, confirmed and its mRNA in SiHa with rmBD2 could be detected at 4 weeks, 6 weeks, 8 weeks, and 10 weeks after cell transfected. A 220 bp segment encoding rmBD2 was amplified by RT-PCR and proved by sequencing. The SiHa/K cells transfected pcDNA3. 1 (+) and SiHa had no expression of rmBD2 protein. The SiHa with rmBD2 expression grew slowly than SiHa/K and SiHa control (P < 0.05).
Conclusion:
The screening for SiHa with rmBD2 expressing stably is successful. The cell growth curve shows that rmBD2 can inhibit the proliferation of tumor cells. The above results establish a solid foundation for further studying the biological properties and the anti-tumor mechanism of beta defensins.
